超声波辐照联合微泡助溶血栓的体外实验研究
发布时间:2018-07-12 10:30
本文选题:超声 + 微泡 ; 参考:《川北医学院》2017年硕士论文
【摘要】:目的:1.模拟体外流动模型,观察超声波辐照联合微泡在血栓溶解中的作用,评估超声波辐照增强血栓溶解效果。2.在体外实验中对比自制脂质微泡(microbubbles,MBs)与Sono Vue的血栓溶解差异,观察不同类型微泡对血栓溶解的影响。3.在体外实验中对比观测37℃与41℃不同环境温度的血栓溶解差异,了解温度对体外血栓溶解的影响。4.在体外实验中对比新鲜血栓与回缩血凝块的血栓溶解差异,了解超声波辐照联合微泡在不同栓龄血栓中的溶解作用。方法:1.脂质微泡的制备:二棕榈酰磷脂酰胆碱(DPPC)、聚乙二醇二硬脂酰磷脂酰乙胺(DSPE-PEG)、甘油、全氟丙烷,通过机械震荡法制备脂质微泡。2.血栓制备:1新鲜血栓:用枸橼酸真空采血管抽取健康志愿者全血,氯化钙溶液促进血液凝固,放入37℃温浴箱温浴3h后,将血栓放入4℃冰箱储存3h。2回缩血凝块:用枸橼酸真空采血管抽取健康志愿者全血,氯化钙溶液促进血液凝固,37℃温浴箱温浴3h后,在4℃冰箱储存7d,7d后取出试管,将半透明的血清从血凝块中分离,以获得稳定的回缩血凝块。3.体外循环装置:由密闭模拟血管管道、恒温水浴箱、微量蠕动泵、药物注入通道组成。整个实验过程密闭管道置于恒温水浴中,循环管道流速控制在4.5ml/s,超声波辐照探头置于血栓垂直正上方5cm处固定。4.实验分组:按超声波辐照、微泡、尿激酶、环境温度以及血栓栓龄的不同组合,随机分为1个对照组、6个实验组,每组10个样本,超声波辐照参数统一设定为频率1MHz,强度2w/cm2,占空比50%。A组:超声波假辐照20min,生理盐水2ml,水浴温度37℃,新鲜血栓。B组:超声波辐照20min,MBs 1ml,生理盐水1ml,水浴温度37℃,新鲜血栓。C组:超声波辐照20min,尿激酶1ml,生理盐水1ml,水浴温度37℃,新鲜血栓。D组:超声波辐照20min,MBs 1ml,尿激酶1ml,水浴温度37℃,新鲜血栓。E组:超声波辐照20min,Sono Vue 1ml,尿激酶1ml,水浴温度37℃,新鲜血栓。F组:超声波辐照20min,MBs 1ml,尿激酶1ml,水浴温度41℃,新鲜血栓。G组:超声波辐照20min,MBs 1ml,尿激酶1ml,水浴温度37℃,回缩血凝块。5.应用西门子ACUSON SC2000超声诊断仪和L9-1超声探头测量狭窄率和血栓面积。观察指标:血栓溶解前后的血栓重量损失率、血栓溶解前后体外循环管道的血栓放置部位管腔狭窄率、血栓溶解前后的血栓最大长轴切面面积减少率。6.统计学处理:七组的测量数据经正态性检验均服从正态分布(P0.05),方差同质性检验,方差齐(P0.05),七组的各变量均为连续性数值变量,各组变量以均值±标准差()表示,七组间整体比较采用单因素方差分析,评价七组血栓溶解效果的整体趋势及差异;两两间比较采用LSD-t分析,进行超声波辐照联合微泡、微泡类型、温度、栓龄分别对血栓溶解作用的分析。均以P0.05为具有统计学意义。所有数据均采用SPSS 21.0统计软件进行分析。结果:1.七组血栓溶解前后重量损失率、面积减少率及管腔狭窄率改变,各组间整体比较,差异具有显著统计学意义(P0.01)。2.A组与B组:血栓溶解前后重量损失率、面积减少率差异具有显著统计学意义(P0.01),重量损失率从3%增加到10%,同比增高233%,面积减少率从2%增加到7%,同比增高250%,管腔狭窄率的比较无显著统计学差异(P0.05)。3.C组与D组:血栓溶解前后重量损失率、面积减少率差异具有显著统计学意义(P0.01),重量损失率从15%增加到16%,同比增高7%,面积减少率从10%增加到14%,同比增高40%,管腔狭窄率的比较无显著统计学差异(P0.05)。4.D组与E组:血栓溶解前后重量损失率、面积减少率及管腔狭窄率差异均具有显著统计学意义(P0.01),重量损失率从16%增加到21%,同比增高31%,面积减少率从14%增加到25%,同比增高79%,管腔狭窄率从6%增加到12%,同比增高100%。5.D组与F组:血栓溶解前后重量损失率、面积减少率及管腔狭窄率差异均具有显著统计学意义(P0.01),重量损失率从16%增加到29%,同比增高81%,面积减少率从14%增加到31%,同比增高121%,管腔狭窄率从6%增加到13%,同比增高117%。6.G组与A组:血栓溶解前后重量损失率、面积减少率及管腔狭窄率无显著统计学差异(P0.05)。7.G组与D组:血栓溶解前后重量损失率、面积减少率及管腔狭窄率差异均具有显著统计学意义(P0.01),重量损失率从2%增加到16%,同比增高700%,面积减少率从2%增加到14%,同比增高600%,管腔狭窄率从3%增加到6%,同比增高200%。结论:1.本实验初步证明,在体外模拟血管血液流动模型中,超声波辐照(f=1MHz,P=2.0w/cm2,占空比50%)联合微泡可促进血栓溶解。2.Sono Vue与自制脂质微泡均能增强血栓溶解,Sono Vue作用更为显著。3.体外血栓溶解中,温度的改变影响新鲜血栓溶解,温度升高可显著增强血栓溶解。4.微泡介导的超声增强尿激酶溶栓,受栓龄的影响明显,不能有效促进回缩血凝块溶解。
[Abstract]:Objective: 1. simulate the flow model in vitro, observe the effect of ultrasonic irradiation combined with microbubbles in thrombolysis and evaluate the difference of thrombolysis between microbubbles, MBs and Sono Vue by ultrasound irradiation enhanced thrombolytic effect in vitro, and observe the effect of different types of microbubbles on thrombolysis in vitro, and the effect of.3. on the thrombolysis in vitro is true in vitro. The difference of thrombolysis between 37 and 41 C was observed and the effect of temperature on thrombolytic dissolution in vitro.4. was compared in vitro to compare the difference of thrombolysis between fresh thrombus and retracted blood clot, and to understand the dissolution of ultrasonic irradiation combined with microbubbles in thrombolytic thrombus. Method: preparation of 1. lipid microbubbles: two Palmiyl phosphatidylcholine (DPPC), polyethylene glycol two stearyl phosphatidylethylamine (DSPE-PEG), glycerin, perfluoropropane, preparation of lipid microbubble.2. thrombus by mechanical shock: 1 fresh thrombus: extract healthy volunteers from healthy volunteers with citric acid vacuum blood vessel, calcium chloride solution to promote blood coagulation, and put blood in a warm bath for 3H in a warm bath box at 37 C The thrombus was placed in the refrigerator at 4 C to store 3h.2 retracted blood clot: the whole blood of healthy volunteers was extracted with citric acid vacuum extraction blood vessel, calcium chloride solution was used to promote blood coagulation, and after a warm bath of 37 centigrade temperature bath for 3h, 7d was stored in the refrigerator at 4 C, and the test tube was taken out after 7d to separate the semitransparent serum from the blood clot to obtain a stable.3. cardiopulmonary bypass device for retracted blood clot: The closed pipe, constant temperature water bath, microperistaltic pump, and drug injection channel were made up. The closed pipe was placed in a constant temperature water bath in the whole experimental process. The flow velocity of the circulating pipe was controlled at 4.5ml/s. The ultrasonic probe was placed at the vertical 5cm above the thrombus and fixed in the.4. experiment. The temperature was irradiated by ultrasound, microbubbles, urokinase, and ambient temperature. The different combinations of thrombus age were randomly divided into 1 control groups, 6 experimental groups, 10 samples in each group, and the ultrasonic irradiation parameters were set as frequency 1MHz, intensity 2w/cm2, duty ratio 50%.A group: ultrasonic false irradiation 20min, physiological saline 2ml, water bath temperature 37, and fresh blood suppository.B group: ultrasonic irradiation 20min, MBs 1ml, normal saline 1ml, water bath temperature Degree 37 C, fresh thrombus.C group: ultrasound irradiation of 20min, urokinase 1ml, physiological saline 1ml, water bath temperature 37,.D group of fresh thrombus: ultrasonic irradiation 20min, MBs 1ml, urokinase 1ml, water bath temperature 37 C, fresh thrombus.E group: ultrasonic irradiation 20min, urokinase, water bath temperature 37, fresh thrombus group: ultrasonic irradiation group: ultrasonic irradiation group 1ml, urokinase 1ml, water bath temperature 41 C, fresh thrombus.G group: ultrasound irradiation of 20min, MBs 1ml, urokinase 1ml, water bath temperature 37, and retracted blood clot.5. using SIEMENS ACUSON SC2000 ultrasound diagnostic instrument and L9-1 ultrasonic probe to measure the stenosis rate and thrombus area. The stenosis rate of the thrombus placement of the extracorporeal circulation pipeline and the reduction rate of the maximum axial section area of thrombus before and after thrombolytic dissolution were.6. statistically treated: the measured data of the seven groups were all subject to normal distribution (P0.05), the variance homogeneity test, Fang Chaqi (P0.05), and the variables of the seven groups were continuous numerical variables, each of the variables By means of mean mean standard deviation (), the overall trend and difference of seven groups of thrombolytic effects were evaluated by single factor variance analysis in the seven groups. The analysis of ultrasonic irradiation combined with microbubbles, microbubbles type, temperature and thrombolytic age in the seven groups was statistically significant for the analysis of thrombolytic effect of thrombolysis in the seven groups. All data were analyzed with SPSS 21 statistical software. Results: 1. seven groups of thrombolytic weight loss rate, area reduction rate and lumen stenosis rate were changed in seven groups. The difference between groups was statistically significant (P0.01).2.A group and B group: the weight loss rate before and after thrombolysis and the difference of area reduction rate were statistically significant Meaning (P0.01), the weight loss rate increased from 3% to 10%, increased by 233%, the area reduction rate increased from 2% to 7%, increased 250% from the same period, and there was no significant difference in the stenosis rate (P0.05).3.C group and D group: the weight loss rate before and after thrombolysis, the difference in area reduction rate had significant statistical significance (P0.01), and the weight loss rate increased from 15% to 15%. The increase was 16%, up 7%, the reduction rate of area increased from 10% to 14%, increased by 40%, and there was no significant statistical difference (P0.05) between group.4.D and E: the weight loss rate, area reduction rate and lumen stenosis rate before and after thrombolysis were statistically significant (P0.01), the weight loss rate increased from 16% to 21%, and increased from year to year. The high 31%, the area reduction rate increased from 14% to 25%, the increase of 79%, the stenosis rate of the lumen increased from 6% to 12%, the increase of the 100%.5.D group and the F group: the difference of the weight loss, the area reduction rate and the stenosis rate of the lumen before and after thrombolysis (P0.01), the weight loss rate from 16% to 29%, the increase of 81% and the reduction rate of the area compared to the same period. From 14% to 31%, the increase was 121%, the stenosis rate of the lumen increased from 6% to 13%, the 117%.6.G group and the A group were higher than the same period. There was no significant difference between the weight loss rate, the area reduction rate and the stenosis rate of the lumen before and after thrombolysis (P0.05) group.7.G and D group: the weight loss rate, the area reduction rate and the narrowing rate of the lumen before and after thrombolysis were both significant With statistical significance (P0.01), the weight loss rate increased from 2% to 16%, increased by 700%, the reduction rate of area increased from 2% to 14%, increased 600% from year to year, the stenosis rate of the lumen increased from 3% to 6%, and increased by 200%. conclusion: 1. in this experiment, in vitro simulated blood flow model, ultrasonic irradiation (f=1MHz, P=2.0w/cm2, duty ratio 50%) combined with f=1MHz, P=2.0w/cm2 and 50%) The microbubbles could promote thrombolytic dissolution of thrombolytic.2.Sono Vue and self-made lipid microbubbles, and the effect of Sono Vue was more significant in thrombolysis in.3. in vitro. The change of temperature affects the dissolution of fresh thrombus. The increase of temperature can significantly enhance thrombolytic dissolved.4. microbubbles mediated ultrasound enhanced urokinase thrombolysis. The effect of thrombolytic age is obvious. The effect promotes the retracted blood clot to dissolve.
【学位授予单位】:川北医学院
【学位级别】:硕士
【学位授予年份】:2017
【分类号】:R54;R743
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